RESUMO
Objective:To explore the protective mechanism of phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway in glucagon like peptide-1 (GLP-1) antagonizing the apoptosis of gestational trophoblasts (HTR-8/SVneo) induced by advanced oxidized protein products (AOPP).Methods:Pregnant trophoblast HTR-8/SVneo were cultured in vitro. The cells were divided into control group, AOPP group, GLP-1 group, AOPP + GLP-1 group and AOPP + GLP-1 + LY294002 group. The control group was cultured in 1640 medium; AOPP group was stimulated with 200 μg/ml AOPP; GLP-1 group was stimulated with 50-100 nmol/L GLP-1 for 1 h; AOPP + GLP-1 group was stimulated with 200 μg/ml AOPP for 48 hours, and then GLP-1 (50-100 nmol/L) was added for 1 hour; In AOPP + GLP-1 + LY294002 group, PI3K inhibitor LY294002 was added on the basis of the intervention of AOPP + GLP-1 group. The expression of PI3K/Akt pathway related protein p-Akt was detected by Western blot. Cell viability was detected by cell counting kit (CCK-8). Enzyme linked immunosorbent assay (ELISA) was used to detect the contents of apoptosis promoter protease caspase-9 and caspase-3, and the contents of apoptosis related proteins Bcl-2, Bax and Cyto-c. Results:After AOPP stimulation, the expression of p-Akt in AOPP group was lower than that in control group ( P<0.05); After 50 and 100 nmol/L GLP-1 intervention, the expression of p-Akt in AOPP + GLP-1 group was significantly higher than that in AOPP group (all P<0.05). After 24 and 48 hours of 100 nmol/L GLP-1 intervention, the expression of p-Akt in AOPP + GLP-1 group was significantly higher than that in AOPP group (all P<0.05). After AOPP stimulation, the cell viability of AOPP group was lower than that of control group ( P<0.05); After GLP-1 intervention, the cell viability of AOPP + GLP-1 group was significantly higher than that of AOPP group ( P<0.05). After adding PI3K inhibitor LY294002, the cell viability of AOPP + GLP-1 + LY294002 group was significantly lower than that of AOPP + GLP-1 group ( P<0.05). The results of ELISA showed that the contents of apoptosis promoter protein caspase-3, caspase-9, apoptosis related protein Bax and Cyto-c in AOPP group were higher than those in control group (all P<0.05), and the content of anti-apoptosis protein Bcl-2 was lower than that in control group ( P<0.05); After GLP-1 intervention, the contents of caspase-3, caspase-9, Bax and Cyto-c in AOPP + GLP-1 group were significantly lower than those in AOPP group ( P<0.05), and the content of anti-apoptosis protein Bcl-2 was higher than that in AOPP group ( P<0.05). After treatment with PI3K inhibitor LY294002, the contents of Bcl-2 in AOPP + GLP-1 + LY294002 group were lower than those in AOPP + GLP-1 group, and the contents of Bax and Cyto-c were higher than those in AOPP + GLP-1 group (all P<0.05). Conclusions:GLP-1 may mediate PI3K / Akt pathway to antagonize the apoptosis of HTR-8/SVneo induced by AOPP.
RESUMO
Objective To explore the effect of a herbalcompound Gehua Jiejue Dizhi Decoction (GJDD) on the liver fat deposition and the expression of PXR, and the mRNA and protein expression of its target genes CYP3A11 and CYP3A25in the liver tissues of mouse models of alcoholic fatty liver.Methods Twenty-nine healthy male C57BL/6J mice were randomly divided into control group (n=5), model group (n=8), high dose GJDD group (n=8)and low dose GJDD group (n=8).The mouse model of alcoholic fatty liver was prepared according to the National Institute on Alcohol Abuse and Alcoholism (NIAAA) method.Then, the mice were treated with the high dose and low dose GJDD for 9 days.Serum glutamic-pyruvic transaminase (AST) and aspartate aminotransferase (AST) were detected by enzyme-linked immunosorbent assay (ELISA).Liver fat deposition was detected by oil red O staining.Real-time RT-PCR and immunohistochemistry were performed to examine the expressions of PXR, CYP3A11 and CYP3A25.Results Compared with the model group, the liver fat deposition in the intervention groups was significantly reduced in a dose-dependent manner, with a significant increase of the expression of PXR and CYP3A25 (P < 0.01).The serum ALT level was significantly reduced in the model group (P < 0.01), while the transcriptional levels of CYP3A11 mRNA in the groups were similar (P ≥ 0.05).Conclusions Gehua Jiejue Dizhi Decoction has obvious therapeutic effect on the AFLD in mice, which may be related to the activation of PXR and its target genes CYP3A25.